Kasni consists of fruit and seed of Cichorium intybus Linn (Fam. Asteraceae).
Kasni contains not less than 0.7 per cent w/w of Chlorogenic acid calculated on the dried basis.
1mm in thickness triangle shaped and has characteristic odour.
ring of pappus scales.
epidermis, with thick cuticle. Numerous rod shaped unequal epicuticluar wax deposited of different size are
present on cuticle. These rod shaped structure are perpendicular to the surface of epidermis. Epidermis is
followed by 6 to 10 layers of thick sclerenchymatous calls. Most of sclerenchymatous calls contains prism if
calcium oxalate crystal, the crystal cell contains single prism.
Determine by thin-layer chromatography (2.4.17), coating the plate with silica gel GF254.
bath. Cool and filter. Reflux the residue with 3 x 25 ml methanol. Combine the extracts and concentrate under
vacuum, to 25 ml with methanol.
Reference solution. Reflux 1gm of Kasni RS with 25 ml of methanol on a water bath. Cool and filter. Reflux the
residue with 3 x 25 ml methanol. Combine the extracts and concentrate under vacuum, to 25 ml with methanol.
plate in air and examine in ultraviolet light at 254 and 365 nm. Spray with a anisaldehyde sulphuric acid reagent.
Heat at 110 º for 10 minutes and examine in the visible light. The chromatographic profile of the test solution is
similar to that of the reference solution.
for 3 hours, cool and filter. Reflux the residue further with methanol till last extract turn colourless, cool & filter.
Combine all the filtrates and concentrates to 10ml
Reference solution. A 0.04 per cent w/v solution of chlorogenic acid RS in methanol.
a stainless steel column 15 cm x 4.6 mm packed with octadecylsilane bonded to porous silica (5µm),
column temperature: 25º,
mobile phase: A. a mixture of 40 volumes of acetonitrile, 60 volumes of buffer solution and adjusted pH
Time Mobile Phase A Mobile Phase B
(in min) (per cent v/v) (per cent v/v)
0 90 10
10 80 20
20 70 30
26 90 10
Inject the reference solution. The test is not valid unless the relative standard deviation for the replicate injections is
not more than 2.0 per cent.
Inject the reference solution and the test solution.
Calculate the content of chlorogenic acid.
Storage. Store protected from heat, moisture and against attack by insects and rodents
(Under UV light at
(Under UV light at
(After spray anisaldehyde
sulphuric acid reagent)
HPLC Chromatogram of Chlorogenic acid
HPLC Chromatogram of Kasni